Cleaved-Caspase 3 (Asp175), p17 Antibody

Product:	Cleaved-Caspase 3 (Asp175), p17 Antibody
Catalog:	AF7022
Description:	Rabbit polyclonal antibody to Cleaved-Caspase 3 (Asp175), p17
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat, Bovine
Prediction:	Pig, Bovine, Horse, Sheep, Rabbit, Dog, Xenopus
Mol.Wt.:	17kDa; 32kD(Calculated).
Uniprot:	P42574
RRID:	AB_2835326

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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50ul	$250	In stock
100ul	$350	In stock
200ul	$450	In stock

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Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Rat,Bovine

Prediction:

Pig(100%), Horse(86%), Sheep(100%), Rabbit(86%), Dog(100%), Xenopus(86%)

Clonality:

Polyclonal

Specificity:

Cleaved-Caspase 3 (Asp175,p17) Antibody detects endogenous levels of fragment of activated Caspase 3 resulting from cleavage adjacent to Asp175.

RRID:

AB_2835326
Cite Format: Affinity Biosciences Cat# AF7022, RRID:AB_2835326.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Stable for 12 months from date of receipt. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

A830040C14Rik; Apopain; CASP-3; CASP3; CASP3_HUMAN; Casp3a; Caspase 3; Caspase 3, apoptosis-related cysteine peptidase; Caspase 3, apoptosis-related cysteine protease; Caspase 3, apoptosis-related cysteine protease a; Caspase-3 subunit p12; CC3; CPP-32; CPP32; CPP32B; Cysteine protease CPP32; EC 3.4.22.56; LICE; mldy; OTTHUMP00000165052; OTTHUMP00000165053; OTTHUMP00000165054; PARP cleavage protease; Procaspase3; protein Yama; SCA 1; SCA-1; SREBP cleavage activity 1; Yama;

Immunogens

Immunogen:

Uniprot:

P42574(CASP3_HUMAN) >>Visit HPA database.

Gene(ID):

CASP3(836)

Expression:

P42574 CASP3_HUMAN:

Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.

Description:

This gene encodes a protein which is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce 2 subunits, large and small, that dimerize to form the active enzyme.

Sequence:

P42574 CASP3_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MENTENSVDSKSIKNLEPKIIHGSESMDSGISLDNSYKMDYPEMGLCIIINNKNFHKSTGMTSRSGTDVDAANLRETFRNLKYEVRNKNDLTREEIVELMRDVSKEDHSKRSSFVCVLLSHGEEGIIFGTNGPVDLKKITNFFRGDRCRSLTGKPKLFIIQACRGTELDCGIETDSGVDDDMACHKIPVEADFLYAYSTAPGYYSWRNSKDGSWFIQSLCAMLKQYADKLEFMHILTRVNRKVATEFESFSFDATFHAKKQIPCIVSMLTKELYFYH

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Pig

100

Bovine

100

Sheep

100

Dog

100

Horse

Xenopus

Rabbit

Chicken

Zebrafish

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P42574 As Substrate

P42574

Site	PTM Type	Enzyme	Source
M1	Acetylation		Uniprot
T4	Phosphorylation		Uniprot
S7	Phosphorylation		Uniprot
S10	Phosphorylation		Uniprot
K11	Acetylation		Uniprot
K11	Ubiquitination		Uniprot
S12	Phosphorylation		Uniprot
K14	Ubiquitination		Uniprot
K19	Ubiquitination		Uniprot
S24	Phosphorylation		Uniprot
S26	Phosphorylation		Uniprot
S29	Phosphorylation		Uniprot
Y41	Phosphorylation		Uniprot
K57	Ubiquitination		Uniprot
S65	Phosphorylation		Uniprot
T67	Phosphorylation		Uniprot
K82	Acetylation		Uniprot
K82	Ubiquitination		Uniprot
K88	Ubiquitination		Uniprot
K105	Ubiquitination		Uniprot
K138	Ubiquitination		Uniprot
S150	Phosphorylation	Q16539 (MAPK14)	Uniprot
T152	Phosphorylation		Uniprot
C163	S-Nitrosylation		Uniprot
T174	Phosphorylation		Uniprot
S176	Phosphorylation		Uniprot
K210	Ubiquitination		Uniprot
K229	Ubiquitination		Uniprot
S249	Phosphorylation		Uniprot
K260	Ubiquitination		Uniprot
T270	Phosphorylation		Uniprot

Research Backgrounds

P42574_CASP3_HUMAN

Function:

Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-|-Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin. Triggers cell adhesion in sympathetic neurons through RET cleavage.

PTMs:

Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.

S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.

Subcellular Location:

Cytoplasm.

Tissue Specificity:

Subunit Structure:

Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 17 kDa (p17) and a 12 kDa (p12) subunit. Interacts with BIRC6/bruce.

Family&Domains:

Belongs to the peptidase C14A family.

Research Fields

· Cellular Processes > Cell growth and death > p53 signaling pathway. (View pathway)

· Cellular Processes > Cell growth and death > Apoptosis. (View pathway)

· Cellular Processes > Cell growth and death > Apoptosis - multiple species. (View pathway)

· Environmental Information Processing > Signal transduction > MAPK signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > TNF signaling pathway. (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).

· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.

· Human Diseases > Neurodegenerative diseases > Parkinson's disease.

· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).

· Human Diseases > Neurodegenerative diseases > Huntington's disease.

· Human Diseases > Infectious diseases: Bacterial > Epithelial cell signaling in Helicobacter pylori infection.

· Human Diseases > Infectious diseases: Bacterial > Pertussis.

· Human Diseases > Infectious diseases: Bacterial > Legionellosis.

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Parasitic > Amoebiasis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.

· Human Diseases > Cancers: Overview > Pathways in cancer. (View pathway)

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Colorectal cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Small cell lung cancer. (View pathway)

· Human Diseases > Cardiovascular diseases > Viral myocarditis.

· Organismal Systems > Immune system > Natural killer cell mediated cytotoxicity. (View pathway)

· Organismal Systems > Immune system > IL-17 signaling pathway. (View pathway)

· Organismal Systems > Nervous system > Serotonergic synapse.

References

1). CircGPR137B/miR-4739/FTO feedback loop suppresses tumorigenesis and metastasis of hepatocellular carcinoma. Molecular Cancer, 2022 (PubMed: 35858900) [IF=37.3]

Application: WB Species: Human Sample: HepG2 and Hep3B cells

Fig. 6 FTO is regulated by circGPR137B/miR-4739 axis in HCC. A Schematic representation of the binding sites between miR-4739 and FTO 3’UTR. B Comparison of the luciferase activity of WT or Mut FTO 3’UTR after treatment with miR-4739 mimics in HepG2 and Hep3B cells. C, D RT-qPCR and western blot analysis of the expression of FTO, p21, p27, cleaved caspase-3/− 9, N-cadherin, E-cadherin and vimentin after the transfection with circGPR137B lentiviruses and (or) miR-4739 mimics in HepG2 and Hep3B cells. E, F, G MTT, colony formation and transwell analysis of the cell proliferation, colony number and cell invasion after the transfection with FTO plasmids and (or) miR-4739 mimics in HepG2 and Hep3B cells. Data are the means ± SEM of three experiments. *P < 0.05, **P < 0.01, ***P < 0.001

2). Ultrasound triggered local sequential reactive oxygen species and nitric oxide release for enhanced cerasomal drug delivery. Chemical Engineering Journal, 2024 [IF=15.1]

3). Upregulation of BCL-2 by acridone derivative through gene promoter i-motif for alleviating liver damage of NAFLD/NASH. NUCLEIC ACIDS RESEARCH, 2020 (PubMed: 32710621) [IF=14.9]

Application: WB Species: human Sample: HepG2

Figure 4. Effect of A22 on anti-apoptosis in 0.5 mM palmitic acid oil (PA) induced cell model. (A) Effect of A22 on cell viability for anti-apoptotic protective effect. (B) Effect of A22 on transcription of BCL-2 and BAX with measurement of mRNA levels. (C) Effect of A22 on protein expressions related with apoptosis (left), which were quantitatively analyzed (right). All the experiments were repeated for three times.

4). Antitumor activity of aumolertinib, a third-generation EGFR tyrosine kinase inhibitor, in non-small-cell lung cancer harboring uncommon EGFR mutations. Acta Pharmaceutica Sinica B, 2023 (PubMed: 37425047) [IF=14.5]

Application: WB Species: Mouse Sample: tumor tissue

Figure 6 Antitumor activity of aumolertinib in the PDX model harboring EGFR H773-V774insNPH mutation. Tumor volume (A), tumor weight (B), average body weights of mice at treatment start (Day 0) and study end (Day 26) (C), and the photograph of tumors (D) in a PDX LU0387 (EGFR H773_N774insNPH) NSCLC tumor model in mice over 26-day treatment with aumolertinib. The vehicle was used as the control. (E, F) Expression of pEGFR, EGFR, pAKT, AKT, pERK, ERK, PCNA, cleaved PARP, PARP, cleaved caspase-3, and caspase-3 in tumor tissue after administration of EGFR-TKIs. Tumors were obtained and lysed at 6 h after the last treatment. Data are presented as the ratio relative to the vehicle group. The data of (A), (B), (C), and (F) are presented as the mean ± SEM (n = 6/group) and compared with the vehicle (∗P < 0.05; ∗∗P < 0.01). PDX, patient-derived xenografts; EGFR, epidermal growth factor receptor; TKIs, tyrosine kinase inhibitors; NPH, H773-V774insNPH.

5). Prenatal exposure to environmentally relevant levels of PBDE-99 leads to testicular dysgenesis with steroidogenesis disorders. JOURNAL OF HAZARDOUS MATERIALS, 2022 (PubMed: 34879533) [IF=13.6]

6). 2,3′,4,4′,5-Pentachlorobiphenyl induces mitochondria-dependent apoptosis mediated by AhR/Cyp1a1 in mouse germ cells. Journal of Hazardous Materials, 2023 (PubMed: 37055962) [IF=13.6]

7). Cyanidin-3-O-glucoside restores spermatogenic dysfunction in cadmium-exposed pubertal mice via histone ubiquitination and mitigating oxidative damage. JOURNAL OF HAZARDOUS MATERIALS, 2020 (PubMed: 31796358) [IF=13.6]

Application: WB Species: Mice Sample: testis

Figure 9 The representative photographs and grayscale analysis of proteins in testis from mice treated for 30 days involved in MAPK signaling pathway. (A) p53 and p-p53. (B) Bax, Bcl-2, and Bad. (C) Total Caspase 3 and Cleaved-Caspase 3. (D) Full PARP and Cleaved-PARP. Comparison between all groups was evaluated through One-way ANOVA. Mean±SD. n=4. **p<0.01, *p<0.05, compared with control group. ##p<0.01, #p<0.05, compared with Cd group. ns, not significant.

8). The role of the apoptosis-related protein BCL-B in the regulation of mitophagy in hepatic stellate cells during the regression of liver fibrosis. EXPERIMENTAL AND MOLECULAR MEDICINE, 2019 (PubMed: 30635551) [IF=12.8]

Application: WB Species: mouse Sample: HSC apoptosis

Fig. 3| Activation of mitophagy induces apoptosis in HSCs. a Mitochondrial DNA (mtDNA) measured by PicoGreen staining in LX2 cells; scale bar,10 μm. b Representative western blots of TOM20 with GAPDH serving as the internal reference. Bar graph represents the mean ± SEM. *P < 0.05 vs.the indicated groups. c TUNEL staining of LX2 cells from the indicated groups; scale bar, 25 μm. Bar graph represents the mean ± SEM. *P < 0.05 vs.the indicated groups. D Annexin V-FITC/PI double-staining and flow cytometry analysis of LX2 cells. Bar graph represents the mean ± SEM. **P < 0.01 vs. the indicated groups. e Representative western blots of cleaved caspase3, cleaved caspase9, collagen I and α-SMA. Bar graph represents the mean± SEM of three different experiments. *P < 0.05 and **P < 0.01 vs. the indicated groups

9). HSP70 attenuates compression-induced apoptosis of nucleus pulposus cells by suppressing mitochondrial fission via upregulating the expression of SIRT3. EXPERIMENTAL AND MOLECULAR MEDICINE, 2022 (PubMed: 35338257) [IF=12.8]

Application: WB Species: Rat Sample: NP cells

Fig. 4 HSP70 inhibited the mitochondrial apoptotic pathway. a, b The MMP of NP cells measured by flow cytometry using JC-1 staining. c, d The production of mtROS in NP cells measured by flow cytometry using MitoSOX Red staining. e, f The production of cellular ROS in NP cells measured by flow cytometry using DCFH-DA staining. g The effects of TRC on ATP production in NP cells. h The MMP and production of mtROS and cellular ROS in NP cells measured by flow cytometry. i The effects of VER on ATP production in NP cells treated with TRC. j The effects of TRC on the expression of cleaved caspase 3, cleaved caspase 9, Bax, and Bcl-2. k The effects of VER on the expression of cleaved caspase 3, cleaved caspase 9, Bax, and Bcl-2 in NP cells treated with TRC. (N = 3, *P < 0.05; **P < 0.01; ***P < 0.001).

Application: IHC Species: Rat Sample: NP cells

10). TFE3, a potential therapeutic target for Spinal Cord Injury via augmenting autophagy flux and alleviating ER stress. Theranostics, 2020 (PubMed: 32802192) [IF=12.4]

Application: WB Species: mice Sample: spinal cord

Figure 2. Inhibition of autophagy activity aggravates ER stress-induced apoptosis after SCI. (A) Western blot analysis of autophagy flux markers (ATG5, Beclin1, VPS34, C-CTSD, SQSTM1/p62, UB and LC3) and ER stress-induced apoptosis markers (CHOP, CASP12, C-CASP12, C-CASP3) in spinal cord lesions from ATG5-/+ mice and ATG5+/+ mice with and without SCI, at Day3 after SCI. (B, C) The levels of the autophagy flux makers from (A) normalized to loading control GAPDH. (D) The expressions of ER stress-induced apoptosis markers from (A) normalized to loading control GAPDH. (E) Image (30×) of spinal cord sections from the indicated groups at Day 3 stained with antibodies against LC3II/NeuN, p62/NeuN, CHOP/NeuN,and CASP3/NeuN, respectively. Scale bar: 25 µm. (F) Quantification of immunofluorescence data from (E) showing the mean number of LC3II in motor neurons at the spinal cord. (G-I) Quantification of immunofluorescence data from (E) showing the mean optical density of p62, CHOP, and CASP3, respectively, in motor neurons of spinal cord. n=6, ns stands for not significant, *P<0.05, **P<0.01.

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Cleaved-Caspase 3 (Asp175), p17 Antibody - #AF7022